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比伐卢定对大鼠背部随意型皮瓣成活的影响及相关机制

Effects and related mechanism of bivalirudin on the survival of random skin flap on the back of rat

摘要:

目的 探讨比伐卢定对大鼠背部随意型皮瓣成活的影响及相关机制. 方法 将30只SD大鼠按随机数字表法分为比伐卢定和生理盐水组,每组15只.在2组大鼠背部制作9 cm×3 cm随意型皮瓣模型.伤后即刻,比伐卢定组大鼠腹腔注射5 mg/mL的比伐卢定注射液0.8 mL/kg,生理盐水组大鼠腹腔注射灭菌生理盐水0.8 mL/kg,每天注射1次,连续注射7d.依据皮瓣血供情况将皮瓣三等分为远端区、中间区和近端区,伤后第1、3、7天,肉眼观察皮瓣各分区大体成活情况.伤后第7天,计算皮瓣成活率;切取皮瓣3个分区中心部位的皮肤组织,HE染色观察这3个分区皮肤组织形态,计算皮瓣中间区微血管密度(MVD);免疫组织化学染色检测皮瓣中间区血管内皮生长因子(VEGF)表达.对数据行t检验. 结果 (1)伤后第1天,2组大鼠皮瓣都出现不同程度的肿胀,主要集中在远端区,但未出现明显的坏死;皮瓣中间区和近端区处于成活状态.伤后第3天,2组大鼠皮瓣坏死集中在中间区;皮瓣近端区仍均处于成活状态;皮瓣远端区均出现大部分坏死,有少许结痂.伤后第7天,2组大鼠皮瓣中间区坏死局部均逐渐融合,比伐卢定组大鼠皮瓣成活面积大于生理盐水组;皮瓣远端区基本坏死;皮瓣近端区基本成活.(2)伤后第7天,比伐卢定组大鼠皮瓣成活率为(64±4)%,明显高于生理盐水组的(45±3)%(t=13.49,P<0.0l).(3)伤后第7天,2组大鼠皮瓣远端区组织形态相似,炎性细胞广泛浸润,组织水肿明显.比伐卢定组大鼠皮瓣中间区出现大量新生血管,伴有新生血管侧支的形成,新生血管基本可见扩张;生理盐水组大鼠皮瓣中间区所形成的新生血管较少,新生血管扩张不明显.2组大鼠皮瓣近端区均可见少量炎性细胞浸润;与生理盐水组相比,比伐卢定组大鼠皮瓣近端区组织水肿程度较轻,血管扩张较多.(4)比伐卢定组大鼠皮瓣中间区MVD为(26±5)个/mm2,明显高于生理盐水组的(18 ±3)个/mm2(t=5.43,P<0.05).(5)比伐卢定组大鼠皮瓣中间区VEGF表达量为6 534±384,明显高于生理盐水组的4659±448(t=12.31,P <0.05). 结论 比伐卢定可以促进大鼠随意型皮瓣成活,其机制可能为减少血栓形成,改善皮瓣血供,增加VEGF表达,促进皮瓣新生血管形成.

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abstracts:

Objective To investigate the effects and related mechanism of bivalirudin on the survival of random skin flap on the back of rat.Methods Thirty SD rats were divided into bivalirudin group and normal saline group according to the random number table,with 15 rats in each group.The random flap model with size of 9 cm ×3 cm was reproduced on the back of rats in two groups.Immediately post injury,rats in bivalirudin group were intraperitoneally injected with 5 mg/mL bivalirudin (0.8 mL/kg),while rats in normal saline group were intraperitoneally injected with normal saline (0.8 mL/kg) once a day.The continuous injection lasted for 7 days.The flap was divided into distal area,middle area and proximal area averagely based on the flap blood supply.On post injury day (PID) 1,3,and 7,the overall survival of each area of flap was observed with naked eyes.On PID 7,the survival rate of flap was calculated,and then the morphology of skin tissue at the center of the three areas of flap was observed by HE staining,the microvessel density (MVD) of the middle area of flap was calculated,and the expression of vascular endothelial growth factor (VEGF) of the middle area of flap was detected with immunohistochemical staining.Data were processed with t test.Results (1) On PID 1,flaps of rats in two groups had different degrees of swelling,mainly concentrated in distal area,but there was no obvious necrosis.The middle area and proximal area of flaps in two groups were survived.On PID 3,the necrosis of flaps of rats in two groups was concentrated in the middle area,while the proximal area of flap was still in survival state,and most distal area of flap was necrosis with a little scab.On PID 7,the necrosis of middle area of flaps of rats in two groups was gradually fused,and the survival area of flap of rats in bivalirudin group was larger than that in normal saline group.The distal area of flap was almost necrotic,and the proximal area of flap was almost survived.(2) On PID 7,the survival rate of flap of rats in bivalirudin group was (64 ± 4) %,significantly higher than that in normal saline group [(45 ±3)%,t =13.49,P <0.01].(3) On PID 7,the histological morphology of distal area of flap of rats in two groups was similar,the inflammatory cells were infiltrated abundantly,and tissue edema was obvious.A large number of new blood vessels appeared in the middle area of flap of rats in bivalirudin group,with the formation of collateral vessels,and basic dilation of new blood vessels was seen.There were fewer new blood vessels appeared in the middle area of flap of rats in normal saline group,and dilation of new blood vessels was not obvious.There was little inflammatory cells infiltration in the proximal area of flap of rats in two groups.Compared with that in normal saline group,tissue edema extent of proximal area of flap of rats in bivalirudin group was less,and expansion was observed in more blood vessels.(4) The MVD of middle area of flap of rats in bivalirudin group was (26 ± 5)/mm2,significantly higher than that in normal saline group [(18±3)/mm2,t =5.43,P <0.05].(5) The expression of VEGF of middle area of flap of rats in bivalirudin group was 6 534 ± 384,significantly higher than that in normal saline group (4 659 ± 448,t =12.31,P < 0.05).Conclusions Bivalirudin can promote the survival of random skin flap in rats,and the mechanisms may include reducing the formation of thrombosis,improving the blood supply of flap,and increasing the expression of VEGF,promoting the formation of new blood vessels.

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作者: 蔡乐益 [1] 王特 [1] 林丁盛 [1] 卢迪 [1]
期刊: 《中华烧伤杂志》2017年33卷4期 228-232页 MEDLINEISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.issn.1009-2587.2017.04.008
发布时间: 2017-05-16
基金项目:
国家自然科学基金 浙江省医药卫生一般研究计划(2016KYB195)National Natural Science Foundation of China General Research Program of Medicine and Health of Zhejiang Province
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